Part:BBa_K4140013

VSVg-Fusogen

Part Description

The Vesicular Stomatitis Virus G (VSV G) protein is a typical type III viral fusion protein that, when present in cell-free environments, forms complexes with plasmid DNA and particles that resemble MLV retroviruses and improves DNA transfection.

Usage

(VSV G) protein is a typical type III viral fusion protein that, considered the second component of SEND platform for RNA delivery we use it to deliver PAH mRNA to specific type of cells(hepatocyte) this will participate in decreasing the side effect and improving dose efficacy.

Characterization of Mutational Landscape

After creating a multiple sequence alignment of the protein sequence and predicting mutational landscapes, the effect of these mutations on the evolutionary fitness of the protein is tested. The prediction of the mutational landscape by saturation mutagenesis of the VSVg-Fusogen protein. The (Y148H) mutation, as depicted in the chart, had the greatest score when compared to other mutations. On the other hand, it's clear that the (A56T) had the least evolutionary fitness for VSVg-Fusogen protein. As displayed in Figure(1)

Figure 1. (shows the mutational landscape of the VSVg-Fusogen protein.)

Literature Characterization

In this study, HECA2 cells were infected with VSVIND at an MOI of 2 and cultured in the presence or absence of ammonium chloride Fig.1. At 24 hpi, the virus released from HECA2 cells was titrated by plaque assay in BHK-21 cells and expressed as PFU per cell. This study showed that ammonium chloride doesn’t affect virus yield.

Figure 1. Treatment of HECA2 cells with NH4Cl

Figure 2. Virus yield in different ammonium chloride concentration

References

1. Roberts, P. C., Kipperman, T., & Compans, R. W. (1999). Vesicular stomatitis virus G protein acquires pH-independent fusion activity during transport in a polarized endometrial cell line. Journal of virology, 73(12), 10447-10457.‏ Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1396
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 1176
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 987
  
• 1000
  COMPATIBLE WITH RFC[1000]